Question

Can you answer and explain it please

Q. You want to PCR amplify the underlined/bold DNA fragment. Design two primers for this job. The size of each primer is 18 bp -3 Forward primer: 5'- Reverse primer: 5'-( ATGACTATAGGGAACACTACGAGTGTGACAACGGTACGTAAC GCGGCGATCGAGCG ACCGAA GCTACTCCGCCTTTAGCTCTCGACAAT ATCGACGCCGCTA CGGATGTGTTTAGGCCTGGT GA TGCTGG AACCCTTTA TTCGCCGAGGAAG ATGTTATCAGCGGGTCTATCAGCATCA CGATCATCGTGAGCT GGTTCTTGGCTATTTCGGAGCAGGT GTCGAATCCGATGCT ITTGTGGGTGAGATGGGTTTATTCA GTGAAGTC ATCTTACGTACACGGA ACTCAGTGTGAATTGG AAGGC AGTTTAGCGAGTGATGCGCCGCGGATTTTGTATGCGATAGGTAT GCAGTTATCTAAGCGTTTATTGCTTACCAC

Answer 1

Select 2 primers from bold/underlining DNA :

Forward primer : 5' ( ACTACGAGTGTGACAACG ) 3'

Reverse Primer : 5' ( CCACTTCGCGCGAGCATT ) 3'

- Forwerd primer is binds to the first bold section.

- A forward primer is a 18 bp single strand of DNA.

- The most important thing about a primer is making sure it will bind complimentarily to your target DNA sequence.

- If done correctly and the length is good, the primer should bind in one spot only: where you want it to.

- It can be used for sequencing (to find out the sequence of things that come after it in ) direction, or 5' to 3'. Most likely, a forward primer is used for PCR.

- However, in order to do PCR, you need a reverse primer.

- Reverse primer selecte from last row of bold DNA fragment.

- Notice that this doesn't match the sequence I have provided.

- This is because DNA is double stranded, so if you want to prime in the opposite direction, you need to compliment the bases, and read in the opposite direction.

5' ATGC 3' becomes:

| | | |

3' TACG 5'

Notice how they are complimentary? Now, read it from 5' to 3', and you will have your reverse primer.